1. Field of the Invention
The present invention relates to novel phenyldiazirine compounds, biotinylated phenyldiazirine compounds and saccharide-linked biotinylated phenyldiazirine compounds. It further relates to a photoreactive labeling reagent consisting of the biotinylated phenyldiazirine compound expected to be used as a probe for analysis of structure of biosubstances and also to a photoaffinity labeling reagent consisting of the saccharide-linked biotinylated phenyldiazirine compound.
2. Description of the Related Art
Photoaffinity labeling reagents have been known as useful probes which are able to analyze the interacting moiety of pharmaceuticals or ligand with protein. An azide group has been known already as a photoreactive group for labeling the protein by means of a photoreaction and, as a better photoreactive group, a diazirine group has been receiving attention. Introduction of a diazirine group into a ligand derived from biosubstances and pharmaceuticals is usually carried out by means of an amide bond or an ester bond and, for further object of a microdetection, it is converted to a radioactive derivative.
Quite recently, phenyldiazirine derivatives having a biotin residue via a spacer compound in a molecule have been developed as the substances which are able to achieve both affinity purification and microdetection of protein and, for example, it has been reported that non-radioactive photoaffinity labeling reagent can be synthesized by linking a saccharide compound as a ligand derived from biosubstances with a carboxyl group of the diazirine derivative (cf. Yuki Gosei Kagaku Kyokaishi, Vol. 56, No. 7, pages 581-590 (1998); Pharmacia, Vol. 34, No. 8, pages 772-776 (1998); Biochem. J., 330, 1209-1215 (1998); J. Am, Chem. Soc. Vol. 120, pages 453-454 (1998); Bioorganic & Medicinal Chemistry, Vol. 2, No. 12, pages 1367-1373 (1994); Carbohydrate Research, Vol. 294, pages 95-108 (1996); Reviews on Heteroatom Chem., Vol. 14, pages 213-243 (1996); and Chem. Pharm. Bull., Vol. 47, pages 667-671 (1999)). However, according to such a method, it is necessary in linking a saccharide compound to the diazirine compound that an amino group is previously introduced into the saccharide compound and that a carboxyl group of the diazirine compound is made to react therewith in the presence of a condensing agent such as dicyclohexylcarbodiimide (DCC) and, therefore, the operation is troublesome.
On the other hand, as a method for a specific modification of a biosubstance, utilization of an oxyamino group has been receiving attention although an attempt of introducing the oxyamino group into a photoreactive compound and to link to a saccharide compound has not been carried out yet.